Enzolution(R) eIF4A1 ATPase FI Assay System

The Enzolution EIF4A1 ATPase Assay System is intended for use with the Transcreener(R) ADP Assay Kits to measure the RNA-dependent ATPase activity of EIF4A1 (eukaryotic initiation factor 4A-I), a member of the DEAD-box family of RNA helicase family (DDX2A). EIF4A1 functions as a core component of the EIF4F translation initiation complex in eukaryotic cells and is upregulated in diverse cancers. ADP formation by EIF4A1 can be easily measured with the Transcreener ADP2 assay, a far-red, competitive fluorescence assay that enables single addition, mix-and-read detection in a continuous or endpoint format. The assay has been optimized and extensively validated for high throughput screening (HTS) and inhibitor dose response measurements using most multimode plate readers. The Enzolution EIF4A1 ATPase Assay System provides all reagents required to screen and profile EIF4A1 inhibitors, including purified, full-length human EIF4A1 and yeast RNA, when used with the Transcreener ADP2 Assay Kits, which are available with FP, FI and TR-FRET readouts. The protocol is configured for 384-well plates, use of different multi-well plate formats will require adjustment of reagents concentrations utilized in the assay. Key Applications:  Screening for EIF4A1 inhibitors (or activators)  Generating dose response curves and IC50 values for EIF4A1 inhibitors  Kinetic and mechanistic analyses Protein Function: ATP-dependent RNA helicase which is a subunit of the eIF4F complex involved in cap recognition and is required for mRNA binding to ribosome (PubMed:20156963). In the current model of translation initiation, eIF4A unwinds RNA secondary structures in the 5'-UTR of mRNAs which is necessary to allow efficient binding of the small ribosomal subunit, and subsequent scanning for the initiator codon. As a result, promotes cell proliferation and growth (PubMed:20156963) [The Uniprot Consortium]